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Method for determining activity of 5'-nucleotidase, and 5'-nucleotidase detection kit

A detection kit and nucleotidase technology, which is applied in the field of medical inspection and determination, can solve the problems of slow reaction speed, many types of raw materials used, and high cost

Inactive Publication Date: 2019-01-01
浙江亚培生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the fifth generation method has many disadvantages such as many reaction steps, slow reaction speed, many types of raw materials, and high cost.

Method used

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  • Method for determining activity of 5'-nucleotidase, and 5'-nucleotidase detection kit
  • Method for determining activity of 5'-nucleotidase, and 5'-nucleotidase detection kit
  • Method for determining activity of 5'-nucleotidase, and 5'-nucleotidase detection kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Reagent one:

[0033]

[0034] Reagent two:

[0035]

[0036] The ratio of reagent 1 to reagent 2 is 2:1, the rate method, positive reaction, the wavelength is 340nm, the ratio of the tested sample, reagent 1 and reagent 2 is 8:180:90, the temperature is 37°C, and the serum reacts with reagent 1 for 5 minutes. Add reagent two, delay time for 2 minutes, and detect for 2 minutes.

Embodiment 2

[0038] Reagent one:

[0039]

[0040] Reagent two:

[0041]

[0042] The ratio of reagent 1 to reagent 2 is 4:1, the rate method, positive reaction, the wavelength is 340nm, the volume ratio of the tested sample, reagent 1 and reagent 2 is 6:200:50, the temperature is 37°C, serum and reagent 1 The reaction was carried out for 5 minutes, reagent II was added, the delay time was 2 minutes, and the detection was performed for 2 minutes.

Embodiment 3

[0044] Reagent one:

[0045]

[0046]

[0047] Reagent two

[0048]

[0049]The ratio of reagent 1 to reagent 2 is 3:1, the rate method, positive reaction, the wavelength is 340nm, the volume ratio of the tested sample, reagent 1 and reagent 2 is 8:180:60, the temperature is 37 ℃, serum and reagent 1 The reaction was carried out for 5 minutes, reagent II was added, the delay time was 2 minutes, and the detection was performed for 2 minutes.

[0050] Commercially available 5'-nucleotidase was detected using this detection kit, and the results were

[0051] theoretical value

0

42.5

85

127.5

170

221.5

255

279.5

340nm

Measured value

0.42

44.2

92.8

137.7

181.6

219.6

253.1

290.2

328.7

[0052] The line performance reaches more than 300U / L.

[0053] Using the same clinical serum as the test sample, the test was repeated 20 times, and the results were as follows:

[0054]

[0055] SD=0.8256, ...

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PUM

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Abstract

The invention relates to a method for determining the activity of 5'-nucleotidase, further to a 5'-nucleotidase detection kit, and belongs to the technical field of medical test determination. According to the method, 5-inosine monophosphate used as a substrate is converted and dehydrogenated by using purine nucleoside phosphorylase and xanthine dehydrogenase as tool enzymes, oxidized coenzyme isreduced to form a reduced coenzyme, and the enzyme reaction rate is calculated by detecting the generation amount of the reduced coenzyme at a wavelength of 340 nm. Compared to the method in the priorart, the method of the present invention has advantages of less reaction steps, less kinds of raw materials, strong alkaline-phosphatase resistance, stable reagent performance, wide linear range, high sensitivity and good accuracy.

Description

technical field [0001] The invention relates to a method for measuring the activity of 5'-nucleotidase, and also relates to a 5'-nucleotidase detection kit, which belongs to the technical field of medical examination and measurement. Background technique [0002] 5'-nucleotidase is a group of phosphohydrolases with different biochemical properties from alkaline phosphatase, mainly distributed in the heart, liver, brain, muscle, kidney and lung, etc., and distributed in the bile ducts, hepatic sinusoids in the liver and Kupffer cells, which are most abundant in liver cytoplasm and metabolized by the liver. The increased activity of 5'-nucleotidase in blood is mainly seen in hepatobiliary diseases, liver cancer, hepatitis and liver cirrhosis, especially liver cancer, often accompanied by alpha-fetoprotein, carcinoembryonic antigen and alpha-fucosidase and other liver diseases. Simultaneous detection of functional items, comprehensive analysis of hepatobiliary diseases. [00...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/42C12Q1/32C12Q1/48
CPCC12Q1/32C12Q1/42C12Q1/48G01N2333/9029G01N2333/91142G01N2333/916
Inventor 李丰盛陈壮永
Owner 浙江亚培生物技术有限公司
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